This work demonstrated a pioneer work in the pre-treatment of rice straw by phosphoric acid (H3PO4) for succinate production. The optimized pre-treatment condition of rice straw was at 121 C for 30 min with 2 N H3PO4. With this condition, total sugar concentration of 31.2 g/L with the highest hemicellulose saccharification yield of 94% was obtained. The physicochemical analysis of the pre-treated rice straw showed significant changes in its structure thus enhancing enzymatic saccharification. Succinate concentrations of 78.5 and 63.8 g/ L were produced from hydrolysate liquor (L) and solid fraction (S) of the pre-treated rice straw respectively, with a comparable yield of 86% by E. coli AS1600a. Use of a combined L + S fraction in simultaneous saccharification and fermentation (LS + SSF) further improved succinate production at a concentration and yield of 85.6 g/L and 90% respectively. The results suggested that H3PO4 pre-treated rice straw may be utilized for economical succinate production by E. coli AS1600a.